解毒通络调肝方对2型糖尿病大鼠肝脏组织IRE1相关蛋白表达的影响

Effect of Jiedu Tongluo Tiaogan prescription on expression of IRE1 related protein in liver tissue of type 2 diabetic rats

目的探讨解毒通络调肝方对2型糖尿病(T2DM)大鼠肝脏组织中肌醇依赖酶(IRE)1及其磷酸化蛋白表达的影响。方法选择88只9周龄健康、雄性ZDF大鼠为研究对象,用Purina#5008高脂饲料饲养建立糖尿病大鼠模型,将成模大鼠随机分为模型组,西药组,中药低、中、高剂量组,各给药组分别给予相应的药物灌胃;10只9周龄雄性ZL大鼠以普通饲料饲养作为空白对照组,空白对照组及模型组用等体积蒸馏水灌胃。灌胃饲养至第17周处死大鼠,无菌取肝脏组织,用于Western印迹检测内质网应激(ERS)相关蛋白IRE1α及磷酸化(p)-IRE1α的表达。结果与空白对照组比较,其余各组肝脏IRE1α、pIRE1α蛋白表达显著增多(P<0.01);与模型组比较,中药高剂量组IRE1α蛋白的表达水平减少最为显著(P<0.01),西药组IRE1α蛋白的表达水平虽有减少,但与模型组相比无明显意义(P>0.05);中药高剂量组p-IRE1α蛋白的表达与其他治疗组比较减少明显(P<0.01),西药组下降水平与中药中剂量组相当(P>0.05)。结论解毒通络调肝方防治T2DM的机制之一可能是通过下调IRE1α、p-IRE1α蛋白表达抑制ERS相关通路,从而改善胰岛素抵抗及减少细胞凋亡。

Objective To investigate the effects of Jiedu Tongluo Tiaogan prescription on the expressions of IRE1 and its phosphorylated protein in liver tissue of ZDF rat model of type 2 diabetes.Methods Eighty-eight healthy 9-week-old male ZDF rats were selected as experimental subjects.Diabetic rat models were established using Purina#5008 high-fat diet.The rats were randomly divided into model group and Western medicine group,Jiedu Tongluo Tiaogan prescription low-dose,middle-dose,high-dose groups.The drug-administered groups were given the corresponding drugs.Ten 9-week-old male ZL rats were fed with normal diet as blank control group,and the blank control and model groups were intragastrically treated with equal volume of deionized water.The rats were sacrificed at the 17 th week.The expressions of ERS related proteins IRE1αand p-IRE1αwere detected by Western blot.Results The expressions of IRE1αand p-IRE1αproteins in the liver of each group were increased compared with those of blank control group(P<0.01).Compared with that of model group,the expression level of IRE1αprotein in high-dose group of Jiedu Tongluo Tiaogan prescription was decreased most significantly(P<0.01).Although the expression level of IRE1αprotein in Western medicine group was decreased,it had no significant effect compared with that of model group(P>0.05).The expression of p-IRE1αprotein in Jiedu Tongluo Tiaogan prescription high-dose group was significantly lower than that in other treatment groups(P<0.01),and the decrease in Western medicine group was equivalent to that in Jiedu Tongluo Tiaogan prescription group(P>0.05).Conclusions One of the mechanisms of Jiedu Tongluo Tiaogan recipe in preventing and treating type 2 diabetes might be to inhibit ERS related pathways by down-regulating IRE1αand p-IRE1αprotein expressions,thereby improving insulin resistance and reducing apoptosis.